Lab Rapid HBsAg
: 716
Insert
Intended Use . System for qualitative, rapid detection of hepatitis B
virus surface antigen (HBsAg) in serum, plasma and human whole blood
samples.
Professional use.
[For in vitro diagnostic use only].
Principle . The system consists of a membrane on which anti-HBsAg
antibodies have been immobilized in the test region. When carrying out
the test, the sample is placed to react with the conjugate that contains
particles linked to anti-HBsAg antibodies. The conjugate complexes with
the HBsAg present in the sample. After adding the buffer, the antigen-
conjugate complex migrates across the membrane and finds the test
region in which the anti-HBsAg antibody is immobilized. Thus, a colored
line is formed indicating the presence of HBsAg antigen in the sample. The
absence of a test line indicates a negative result as long as the control line,
used as control of the procedure, appears in the test.
Summar y . The Lab Rapid HBsAg system is an
immunochromatographic method that allows the detection of hepatitis B
virus surface antigen (HBsAg) through a simple, fast and easy-to-
interpret procedure in serum, plasma or whole blood samples. The use of
monoclonal and polyclonal anti-HBsAg antibodies guarantee the high
sensitivity and specificity of the product.
Methodology . Immunochromatography.
Reagents
1. Reaction plates or strips - Store between 2 - 30°C.
Do not freeze. It contains anti-HBsAg1 particle conjugate, anti-HBsAg
antibody 2 and control antibody applied or immobilized in the membrane.
2. Buffer - Store between 2 and 30°C.
Do not freeze. It contains phosphate buffer, sodium chloride, high
molecular weight polypeptide and < 0.1% isothiazolinone.
Unopened reagents, when stored under the indicated conditions, are
stable until the expiration date printed on the label. The Buffer, once
opened, is stable for 2 months. While handling, reagents are subject to
chemical and microbial contamination that can cause reduced stability.
Precautions and warnings
The product should never be frozen or exposed to a temperature above
30°C.
Avoid exposure of reaction plates or strips to ambient moisture.
The usual safety precautions should be used when handling reagents and
samples.
01 English - Ref.: 716
Ref.
The sample volume informed in the ‘‘Procedure’’ section of this
instruction for use must be strictly followed. Changes in the use of this
volume can result in erroneous results.
Since no known test can guarantee that blood samples do not transmit
infections, they should all be considered as potentially infectious. Thus,
when handling them, biosafety rules must be followed.
The Buffer contains isothiazolinone which is toxic. Special care must be
taken to avoid ingestion and, in case of eye contact, wash your eyes
immediately with abundant water and seek medical assistance.
To dispose reagents and biological material, we suggest applying local,
state or federal protection standards.
Required material and not provided
1. Chronometer.
Samples
Serum, plasma (EDTA, citrate, oxalate, heparin) or whole blood (EDTA,
citrate, oxalate, heparin).
The analyte is stable for 3 days at 2-8°C. Store the serum or plasma
sample at temperature equal to or lower than minus 20°C for up to 6
months in a hermetically closed container to prevent evaporation.3
The whole blood sample must not be frozen.
Ensure that the samples are thawed and homogenized before use.
Suspended particles must be removed by centrifugation.
Samples must not be inactivated by heat as they may produce incorrect
results.
Do not use samples with signs of contamination or frozen and thawed
samples repeatedly.
Do not use hemolyzed and/or lipemic samples.
A Standard Operating Procedure (SOP) should be created to establish
adequate procedures for sample collection, preparation and storage. We
emphasize that the errors due to the sample can be much larger than the
errors that occurred during the analytical procedure.
Interferences
No false positive results were observed in samples positive for anti-HAV,
anti-HCV, anti-HEV, anti-HIV, Syphilis, Toxoplasmosis, Rubella, CMV,
H. pylori, Rheumatoid factor, HAMA and MONO.
No interference was observed for samples containing hemoglobin up to
2000 mg/dL, bilirubin up to 1000 mg/dL and albumin up to 2000 mg/dL.
 Procedure
Samples and product must be at room temperature at the time of testing.
Reaction plates
1. Remove the reaction plate from the protective envelope, identify it
properly and place it on a horizontal surface.
2. For serum or plasma, with the aid of a pipette, add 3 drops (0.075 mL)
to the sample well (S); for whole blood, with the aid of a pipette, add 3
drops (0.075 mL) and 1 drop (0.04 mL) of the Buffer to the sample well
(S).
3. Perform the reading of the results between 15 and 30 minutes. Do not
do the reading after 30 minutes.
Reaction strips
1. Place the support card (Test Card) on a horizontal surface and properly
identify it.
2. Remove the film from the adhesive part by pulling the part indicated as
‘‘Peel here’’.
3. Remove the reaction strip from the protective envelope and place it on
the support card.
4. For serum or plasma, with the aid of a pipette, add 3 drops (0.075 mL)
to the sample site; for whole blood, with the aid of a pipette, add 3 drops
(0.075 mL) and 1 drop (0.04 mL) of the Buffer at the sample site.
5. Perform the reading of the results between 15 and 30 minutes. Do not
do the reading after 30 minutes.

3 drops of serum or plasma 3 drops of serum or plasma

Paciente: - - - - - - - -

TestCard
S

HBsAg
C
T

MAX
MAX HBsAg
Specimen

C
T
pad

3 drops of whole blood 3 drops of whole blood

1 drop of buffer 1 drop of buffer

r
ffe
r
ffe

Bu
Bu

Paciente: - - - - - - - -

TestCard
S

HBsAg
C
T

MAX
MAX HBsAg
Specimen

C
T
pad

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INTERPRETING THE RESULTS
Positive
Formation of a colored line in the control region (C) and in the test region (T).

Negative
Formation of a colored line in the control region (C) and absence of line in the test region (T).

Samples that present negative result at 15 minutes must have the result confirmed at 30 minutes.

Invalid
The absence of a line in the control region (C) indicates procedural error or system deterioration. Review the procedure and repeat the test with a new plate or
reaction strip.

Reaction Plates

HBsAg

C C C C C C
G

T T T T T T

Positive Negative Invalid

Reaction Strips
HBsAg

HBsAg

HBsAg

HBsAg

HBsAg

HBsAg

C C C C C C
G

T T T T T T
MAX
MAX

MAX
MAX

MAX
MAX

MAX
MAX

MAX
MAX

MAX
MAX

Positive Negative Invalid

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As it is a screening test, any sample that shows a positive result in a rapid
test must also be analyzed by complementary methodology such as
molecular tests or immunoassay for detection of total anti-HBc,
according to the flowcharts described in the Technical Manual for
Diagnosis of Viral Hepatitis in Adults and Children approved by SVS/MS
Ordinance no. 25, dated December 1, 2015.4
This product is a screening test, so a negative result does not rule out the
possibility of HBV infection. If the suspicion of HBV infection persists, a
new sample should be tested according to the recommendations of the
Technical Manual for Diagnosis of Viral Hepatitis approved by Ordinance
nº 25, dated December 1th, 2015.4 For interpretation of the test result,
other clinical information must be taken into account.
Internal Quality Control5 . The laboratory must maintain an
internal quality control program that clearly defines the applicable
regulations, objectives, procedures, criteria for quality specifications and
tolerance limits, corrective actions and record of activities.
The formation of a colored line in the control region is indicative of
adequate performance of the procedure. To ensure that the system has
not been adversely affected and maintains the established performance
levels, we suggest associating a quality control system using, daily,
known samples, one positive and one negative.
Performance characteristics7
Comparative Studies . Comparison studies have been conducted
using 560 positive and negative samples tested with the
Lab Rapid HBsAg - Labtest system and and with a commercially available
ELISA methodology product (comparative method), by obtaining the
results presented in the following tables.
Lab Rapid HBsAg - Labtest
Comparative Method Positive Negative
Positive 149 1 121 p.: il.
Negative 1 409
Relative sensitivity: 99.3%
Relative Specificity: 99.8%
Efficiency: 99.6%
Kappa Index: 0.99
The Kappa Index greater than 0.80 indicates an excellent concordance6
between the methods, showing that the Lab Rapid HBsAg - Labtest
system is substantially equivalent to the comparative method.
Repeatability - Intra-assay precision . The intra-assay
inaccuracy was verified through the evaluation of three replicates of six
samples, of which five were positive with concentrations of 1, 2, 5, 12 and
20 ng/mL and one negative. Negative and positive results found showed a
perfect agreement with the expected results.
Reproducibility - Interassay precision . The total inaccuracy
was verified through the evaluation of ten replicates of six serum samples
and six plasma samples, of which five were positive with concentrations
of 1, 2, 5, 12 and 20 ng/mL and one negative. The tests were performed
for ten days. The results found showed a perfect agreement with the
expected results.
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Pro-zone Effect . The product does not present pro-zone effect in
samples positive for HBsAg with concentration up to 500 ng/mL or
1000 IU/mL.
Methodological sensitivity . The method is capable of detecting
quantities greater than or equal to 1 ng/mL or 2 IU/mL.
The time of execution of the test can interfere in the concentration of
HBsAg present in the sample, such that the methodological sensitivity of
the product Lab Rapid HBsAg - Ref. 716 could be lower.
Clinical significance . Viral hepatitis are systemic diseases that
mainly involve the liver and have the hepatitis B virus (HBV) as one of the
causing agents. HBV belongs to the Hepadnaviridae family and is capable
of infecting different animals.1 The antigen found on the HBV surface is
called HBsAg. It was discovered in 1966 and was called Australia or Au
antigen.2 HBsAg is one of the five serological markers of the HBV infection
and its presence in the sample indicates active infection, which may be
acute or chronic. In a typical HBV infection, HBsAg will be detected
between 2 to 4 weeks before ALT / GTP levels become abnormal and 3 to 5
weeks before the appearance of symptoms or jaundice. There are four
main subtypes for HBsAg: adw, ayw, adr and ayr and due to the
heterogeneity of the antigen there are more than 10 serotypes for HBV.
HBV is transmitted parenterally and, mainly, sexually, which classifies
hepatitis B as an STI. Vertical transmission (from mother to child) also
occurs in regions of high endemicity.1
References
1. Brazil. Ministry of Health. Secretariat of Health Surveillance. Sexually
Transmitted, HIV/Aids and Viral Hepatitis Infection Surveillance,
Prevention and Control Department. Technical Manual for Diagnosis of
Viral Hepatitis / Ministry of Health, Secretariat of Health Surveillance,
Sexually Transmitted, HIV/Aids and Viral Hepatitis Infection Surveillance,
Prevention and Control Department. - Brasília: Ministry of Health, 2016.
2. Blumberg, B.S. The Discovery of Australian Antigen and its relation to
viral hepatitis.Vitro.1971; 7: 223.
3. Recommendations of the Brazilian Clinical Medicine / Laboratory
Medicine Society (SBPC/ML). Collection and preparation of biological
samples. 2014.
4. Brazil. Ministry of Health. Secretariat of Health Surveillance. SVS/MS
Ordinance no. 25, dated Dececmber 1, 2015. It approves the Technical
Manual for Diagnosis of Viral Hepatitis in Adults and Children, available at
the electronic address www.aids.gov.br, which contains the
recommended flowcharts for different scenarios and situations that suit
the plurality of conditions and the diversity of public and private health
services. Federal Official Gazette (Diario Oficial da União) of the Federative
Republic of Brar zil, December 2, 2015. Available at
<www.saude.gov.br/svs/>.
5. Westgard JO, Barry PL, Hunt MR, Groth T. Clin Chem 1981;27:493-
501.
6. Landis, J. Richard; KOCH, Gary G. The measurement of observer
agreement for categorical data. Biometrics, p. 159-174, 1977
7. Labtest: File on Data
Presentation Labtest Diagnóstica S.A.
CNPJ: 16.516.296 / 0001 - 38
Product Reference Content
Av. Paulo Ferreira da Costa, 600 - Vista Alegre - CEP 33.240-152
Reaction Plate 20 Units Lagoa Santa . Minas Gerais Brasil - www.labtest.com.br
716C-20
Lab Rapid Buffer 1 X 3.0 mL Customer Service e-mail: customerservice@labtest.com.br
HBsAg Reaction Strips 20 Units
716S-20
Buffer 1 X 3.0 mL
Edition: November, 2020 Copyright by Labtest Diagnóstica S.A.
For information on other commercial presentations, please visit Revision: - Reproduction under previous autorization
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Customer information

[Warranty Conditions]

Labtest Diagnóstica guarantees the product performance, within the

specifications, until the expiration date indicated on the labels, provided
that the care for use and storage indicated on the labels and in these
instructions are correctly followed.

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